JPWO2019107440A1 - コレステロール合成促進剤 - Google Patents
コレステロール合成促進剤 Download PDFInfo
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- JPWO2019107440A1 JPWO2019107440A1 JP2019557285A JP2019557285A JPWO2019107440A1 JP WO2019107440 A1 JPWO2019107440 A1 JP WO2019107440A1 JP 2019557285 A JP2019557285 A JP 2019557285A JP 2019557285 A JP2019557285 A JP 2019557285A JP WO2019107440 A1 JPWO2019107440 A1 JP WO2019107440A1
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- Prior art keywords
- differentiation
- cells
- extract
- dhcr7
- epidermal
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Abstract
Description
[1]アセンヤク抽出物、2−メタクリロイルオキシエチルホスホリルコリン基を有するコポリマー、酢酸レチノール、及びシカクマメ抽出物からなる群から選択される少なくとも1種類の成分を含むコレステロール合成促進剤。
[2]1に記載のコレステロール合成促進剤を含む、表皮細胞の分化を促進するための組成物。
[3]表皮細胞の分化を促進する薬剤をスクリーニングする方法であって、候補薬剤を培養細胞に作用させた場合に、該細胞のDHCR7遺伝子の発現を亢進させる薬剤を表皮細胞の分化を促進する薬剤として選定することを特徴とする方法。
[4]前記培養細胞がヒト表皮角化細胞である、3に記載の方法。
[5]表皮細胞の分化を促進するための組成物の調製における、アセンヤク抽出物、2−メタクリロイルオキシエチルホスホリルコリン基を有するコポリマー、酢酸レチノール、及びシカクマメ抽出物からなる群から選択される少なくとも1種類の成分の使用。
[6]DHCR7遺伝子の発現を亢進させる薬剤を含む、表皮細胞の分化を促進するための組成物。
[7]表皮細胞の分化を促進するための美容的又は治療的な方法であって、DHCR7遺伝子の発現を亢進させる薬剤を対象に適用することを含む方法。
[8]表皮細胞の分化を促進するための組成物の調製における、DHCR7遺伝子の発現を亢進させる薬剤の使用。
フォワードプライマー:5'-AGGGGAAGGTGGGCGCAGGAC-3'(配列番号1)
リバースプライマー: 5'-TTGGGCCCTCCAGCCTCTTGC-3'(配列番号2)
二人の被験者の上腕前側部にW−PRP施術を施し、施術前、施術2週間後、施術3か月後に5mmパンチにより皮膚組織の採取を行った。それぞれの試料を液体窒素中でクライオプレスにより破砕し、Isogen(ニッポンジーン社)で細胞内のRNAを抽出し、マニュアル(Agilent 社)に従い、ラベル化プローブを合成し、約4万個の遺伝子がスポットされた、DNAマイクロアレイ(one-color, whole mouse 44K、Agilent社)上で、ハイブリダイゼーションを行った。得られたシグナルはスキャナーAgilent dual-laser Microarray scanner G2565AA を用いて検出し、解析ソフトFeature Extraction Software 9.1 (Agilent 社)を用いて数値化し遺伝子発現指標値とした。
図1に示されるとおり、W−PRP施術により、DHCR7の発現が亢進することが判明した。
ヒト表皮角化細胞をCnT−07培地(CELLnTEC社)で継代培養し、継代4代目の細胞を同培地に懸濁して、コラーゲンコート6穴マルチプレート(旭硝子)に2×105 細胞/ウェルになるように播種し、5%CO2存在下、37℃で細胞が集密に達するまで3〜5日間の培養を行った。CnT−02培地(CELLnTEC社)に交換し、分化誘導を開始した。同時に、ジメチルスルホキシド(DMSO)中に溶解したDHCR7阻害剤(AY−9944)(Sigma-Aldrich社)を最終濃度10μMとなるように添加し、4日間培養した。細胞の状態観察は位相差顕微鏡下で行った。コントロールとしては、DMSOを同量(最終濃度0.1%)添加したサンプルを用いた。
図2の写真に示されるとおり、DHCR7の阻害により、表皮細胞の分化にともなう形態の変化が阻害された。
培養後の細胞からRNA抽出試薬MagNA Pure LC mRNA HSキット(Roche社)と自動核酸抽出装置 MagNA Pure LC 1.0 インスツルメント(Roche社)を用いて、提供されたプロトコールに従ってmRNAの抽出・精製を行った。上述のDHCR7阻害剤(AY−9944)及びコントロールについて、逆転写酵素 Superscript II(Invitrogen社)によりRNAからランダムヘキサマー・プライマーを用いて、Invitrogen社のプロトコールにしたがいcDNAを合成した。合成したcDNAを鋳型に、後述の配列番号1及び2のプライマーペア、反応試薬 QuantiFast SYBR Green RT-PCR キット(Qiagen社)及び反応装置 LightCycler(Roche社)を用いて、分化マーカーであるケラチン10遺伝子の定量リアルタイムPCRを行った。組成条件は Qiagen社のプロトコールに従った。また、PCRの条件は、初期変性95℃で10分、変性95℃で10秒、アニール60℃で10秒、伸長72℃で12秒とした。なお、G3PDHは内部標準として用い、これを用いてコントロール群のmRNA量を補正した。
ケラチン10:
フォワードプライマー:5'-CCATCGATGACCTTAAAAATCAG-3'(配列番号1)
リバースプライマー:5'-GCAGAGCTACCTCATTCTCATACTT-3'(配列番号2)
G3PDH:
フォワードプライマー:5'-GCACCGTCAAGGCTGAGAAC-3'(配列番号3)
リバースプライマー: 5'-ATGGTGGTGAAGACGCCAGT-3'(配列番号4)
図3のグラフに示されるとおり、AY−9944により、表皮細胞の分化マーカーであるケラチン10が顕著に減少した。
上記で調製した分化開始後1日目及び4日目の細胞を200μlのクロロフォルム:イソプロパノール:NP−40(7:11:0.1)で抽出した後、溶媒を蒸散させ、コレステロール/コレステロールエステル定量キット(BioVision社)に含まれるコレステロールアッセイバッファー中に懸濁して測定試料とした。キット付属の96ウェルプレートに試料を添加し、各ウェルに50μlの反応ミックスを加え、37℃で1時間インキュベートした後、プレートリーダーを用い波長570nmの吸光度を測定した。キット付属の標準品を用いて検量線を作成し、サンプル中のコレステロール濃度を定量した。
図4Aに示されるとおり、AY−9944により表皮細胞に含有されるコレステロール量は減少した。
上記で調製した分化開始後1日目の細胞培養液をalamarBlue(R)Cell Viability Reagent(invitrogen社)を10%含有するCnT−02培地(CELLnTEC社)に交換し、5%CO2存在下、37℃で3時間インキュベーションした後、蛍光プレートリーダーを用い励起波長/蛍光波長:540/590nmで計測した。
図4Bに示されるとおり、AY−9944によって細胞増殖に変化は見られなかった。
図4A及び図4Bに示される結果から、実験2におけるケラチン10の減少は細胞死によるものではないこと、及び、DHCR7の阻害により細胞のコレステロール含有量が確かに減少していることが確認された。
ヒト表皮角化細胞をCnT−07培地(CELLnTEC社)で継代培養し、継代3代目の細胞を同培地に懸濁して、コラーゲンコート12穴マルチプレート(旭硝子)に50,000個の割合で播種し、5%CO2存在下、37℃で細胞が集密に達するまで3〜5日間の培養を行った。30品目の候補薬剤を各評価対象サンプルとして、各サンプルを上記濃度となるように添加、又は各評価対象サンプルが溶解している溶媒であるDMSOを同量添加したCnT−07培地に交換した後、更に1日間培養を行った。培養後の細胞からRNA抽出試薬MagNA Pure LC mRNA HSキット(Roche社)と自動核酸抽出装置 MagNA Pure LC 1.0 インスツルメント(Roche社)を用いて、提供されたプロトコールに従ってmRNAの抽出・精製を行った。各サンプルについて、逆転写酵素 Superscript II(Invitrogen社)によりRNAからランダムヘキサマー・プライマーを用いて、Invitrogen社のプロトコールにしたがいcDNAを合成した。合成したcDNAを鋳型に、後述の配列番号1及び2のプライマーペア、反応試薬 QuantiFast SYBR Green RT-PCR キット(Qiagen社)及び反応装置 LightCycler(Roche社)を用いて、DHCR7遺伝子の定量リアルタイムPCRを行った。組成条件はQiagen社のプロトコールに従った。また、PCRの条件は、初期変性95℃で10分、変性95℃で10秒、アニール60℃で10秒、伸長72℃で12秒とした。なお、G3PDHは内部標準として用い、これを用いてコントロール群のmRNA量を補正した。
DHCR7:
フォワードプライマー:5'-AGGGGAAGGTGGGCGCAGGAC-3'(配列番号1)
リバースプライマー: 5'-TTGGGCCCTCCAGCCTCTTGC-3'(配列番号2)
G3PDH:
フォワードプライマー:5'-GCACCGTCAAGGCTGAGAAC-3'(配列番号3)
リバースプライマー: 5'-ATGGTGGTGAAGACGCCAGT-3'(配列番号4)
その結果、アセンヤク抽出物(丸善製薬(株))、MPCコポリマー(日本油脂(株))、酢酸レチノール(BASF社)及びシカクマメ抽出物(一丸ファルコス(株))の4種類の薬剤がDHCR7産生促進効果を有することが判明した(図5)。
ヒト表皮角化細胞をCnT−07培地(CELLnTEC社)で継代培養し、継代3代目の細胞を同培地に懸濁して、コラーゲンコート12穴マルチプレート(旭硝子)に50,000個の割合で播種し、5%CO2存在下、37℃で細胞が集密に達するまで3〜5日間の培養を行った。アセンヤク抽出物(丸善製薬)を0.2%となるように添加、又はコントロールとしてDMSOを同量添加したCnT−07培地に交換した後、更に1日間培養を行った。培養後の細胞からRNA抽出試薬MagNA Pure LC mRNA HSキット(Roche社)と自動核酸抽出装置 MagNA Pure LC 1.0 インスツルメント(Roche社)を用いて、提供されたプロトコールに従ってmRNAの抽出・精製を行った。各サンプルについて、逆転写酵素 Superscript II(Invitrogen社)によりRNAからランダムヘキサマー・プライマーを用いて、Invitrogen社のプロトコールにしたがいcDNAを合成した。合成したcDNAを鋳型に、後述の配列番号1及び2のプライマーペア、反応試薬 QuantiFast SYBR Green RT-PCR キット(Qiagen社)及び反応装置 LightCycler(Roche社)を用いて、分化マーカーであるインボルクリン(involucrin)遺伝W−PRP施術により発現が亢進される遺伝子のマイクロアレイ解析子の定量リアルタイムPCRを行った。組成条件は Qiagen社のプロトコールに従った。また、PCRの条件は、初期変性95℃で10分、変性95℃で10秒、アニール60℃で10秒、伸長72℃で12秒とした。なお、G3PDHは内部標準として用い、これを用いてコントロール群のmRNA量を補正した。
インボルクリン:
フォワードプライマー:5'-CTGCCTCAGCCTTACTGTGA-3'(配列番号5)
リバースプライマー: 5'-TGGGTATTGACTGGAGGAGG-3'(配列番号6)
G3PDH:
フォワードプライマー:5'-GCACCGTCAAGGCTGAGAAC-3'(配列番号3)
リバースプライマー: 5'-ATGGTGGTGAAGACGCCAGT-3'(配列番号4)
図6に示されるとおり、アンヤク抽出物によって、インボルクリンの産生が促進されることが確認された。
Claims (4)
- アセンヤク抽出物、2−メタクリロイルオキシエチルホスホリルコリン基を有するコポリマー、酢酸レチノール、及びシカクマメ抽出物からなる群から選択される少なくとも1種類の成分を含むコレステロール合成促進剤。
- 請求項1に記載のコレステロール合成促進剤を含む、表皮細胞の分化を促進するための組成物。
- 表皮細胞の分化を促進する薬剤をスクリーニングする方法であって、候補薬剤を培養細胞に作用させた場合に、該細胞のDHCR7遺伝子の発現を亢進させる薬剤を表皮細胞の分化を促進する薬剤として選定することを特徴とする方法。
- 前記培養細胞がヒト表皮角化細胞である、請求項3に記載の方法。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005068073A (ja) * | 2003-08-25 | 2005-03-17 | Takashi Fukazawa | 目元用化粧料 |
JP2005281253A (ja) * | 2004-03-30 | 2005-10-13 | Nof Corp | 皮膚化粧料 |
JP2006151834A (ja) * | 2004-11-25 | 2006-06-15 | Nof Corp | 皮膚化粧料 |
US20100203004A1 (en) * | 2007-10-31 | 2010-08-12 | L'oreal | Cosmetic composition containing a tensioning agent and an acrylic polymer |
JP2013177364A (ja) * | 2012-02-10 | 2013-09-09 | Shiseido Co Ltd | 水中油型乳化皮膚化粧料 |
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JP2010143893A (ja) * | 2008-12-22 | 2010-07-01 | Nikko Chemical Co Ltd | セリンプロテアーゼHtrA1及び/又はHtrA2の活性阻害剤 |
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Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2005068073A (ja) * | 2003-08-25 | 2005-03-17 | Takashi Fukazawa | 目元用化粧料 |
JP2005281253A (ja) * | 2004-03-30 | 2005-10-13 | Nof Corp | 皮膚化粧料 |
JP2006151834A (ja) * | 2004-11-25 | 2006-06-15 | Nof Corp | 皮膚化粧料 |
US20100203004A1 (en) * | 2007-10-31 | 2010-08-12 | L'oreal | Cosmetic composition containing a tensioning agent and an acrylic polymer |
JP2013177364A (ja) * | 2012-02-10 | 2013-09-09 | Shiseido Co Ltd | 水中油型乳化皮膚化粧料 |
Non-Patent Citations (2)
Title |
---|
"II. 化粧品の有効成分 2.老化防止剤", 新化粧品ハンドブック, JPN6018051566, 2006, pages 518 - 524, ISSN: 0004876888 * |
鈴木正己: "特集2 ラジオアイソトープ(R.I.)の進歩と応用 ラジオアイソトープの皮膚科学への応用−経皮吸収を", FRAGRANCE JOURNAL NO.28, vol. 6, no. 1, JPN6018051565, 1978, pages 75 - 83, ISSN: 0004876887 * |
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